Graduate School of Life Science, Hokkaido University, Sapporo 060-0810, Japan.
J Mol Histol. 2011 Jun;42(3):251-64. doi: 10.1007/s10735-011-9329-3. Epub 2011 May 5.
Prolyl oligopeptidase (POP) is a serine endopeptidase which selectively digests a -Pro-X- peptide bond. Our previous study showed that POP mRNA was strongly expressed in the spongiotrophoblast of the mouse placenta at E17.5, suggesting its importance in development. To gain more insight into POP's role during gestation, we investigated its expression using different developmental stages of placenta. As a result of in situ hybridization, we found that localization of POP mRNA changed at E12.5. POP mRNA was strongly expressed in the spongiotrophoblast and labyrinth at E10.5 and E11.5 but thereafter only in the spongiotrophoblast. Immunohistochemistry revealed that POP was present in the parietal trophoblast giant cell, the spongiotrophoblast cell, and the labyrinth at E11.5 but the strong expression in the labyrinth was maintained only in the canal-associated and sinusoidal trophoblast giant cells at E16.5 and E18.5. To determine subcellular distribution of the POP protein, we fractionated the placental extract into cytoplasmic, membrane, and nuclear subfractions. By Western blot analysis, POP was detected in the cytoplasmic and membrane fractions but not in the nuclear fraction at E11.5 and E16.5. Interestingly, the cytoplasmic POP exhibited higher enzymatic activity than the membrane-associated type. These data suggest that the cytoplasmic and membrane-associated POP have distinct roles in different types of placental cells.
脯氨酰寡肽酶(POP)是一种丝氨酸内肽酶,它特异性地消化 a-Pro-X-肽键。我们之前的研究表明,POP mRNA 在 E17.5 的鼠胎盘的海绵滋养层中强烈表达,这表明它在发育过程中的重要性。为了更深入地了解 POP 在妊娠期间的作用,我们使用不同发育阶段的胎盘来研究其表达情况。通过原位杂交,我们发现 POP mRNA 的定位在 E12.5 时发生了变化。POP mRNA 在 E10.5 和 E11.5 时在海绵滋养层和胎盘绒毛中强烈表达,但此后仅在海绵滋养层中表达。免疫组织化学显示,POP 存在于壁细胞滋养层巨细胞、海绵滋养层细胞和 E11.5 的胎盘绒毛中,但在 E16.5 和 E18.5 的管相关和窦状滋养层巨细胞中仅维持强烈表达。为了确定 POP 蛋白的亚细胞分布,我们将胎盘提取物分离成细胞质、膜和核亚部分。通过 Western blot 分析,在 E11.5 和 E16.5 时,POP 被检测到在细胞质和膜部分,但不在核部分。有趣的是,细胞质中的 POP 表现出比膜结合型更高的酶活性。这些数据表明,细胞质和膜结合型的 POP 在不同类型的胎盘细胞中具有不同的作用。